Assessment of oxidative stress in vivo

A method to assess oxidative stress in vivo by measuring the amount of free, esterified and glucuronidated forms of isoprostanes (8EPGF2) in a biological sample which contains the isoprostanes is disclosed. The method further includes determining the amount of total isoprostanes present in the sample. This amount is compared with a control sample. The oxidative stress is determined through the comparison wherein the amount of isoprostanes increase in the sample isolated from an organism undergoing oxidative stress compared to the control. Alternatively the method of the present invention provides for only the measurement of the glucuronidated form wherein the amount of glucuronidated isoprostanes increase in the sample isolated from an organism undergoing oxidative stress compared to the control.

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Claims

1. A method to assess oxidative stress in vivo by measuring the amount of free, esterified and glucuronidated forms of isoprostanes in a biological sample containing the isoprostanes and determining a measured amount of total isoprostanes present in the sample, and

comparing the measured amount with a healthy control sample and thereby assessing the oxidative stress through said comparison wherein the total amount of isoprostanes increases in the sample isolated from an organism undergoing oxidative stress in vivo compared to the control.

2. The method of claim 1 wherein the biological sample is selected from the group consisting of plasma, urine, cerebrospinal fluid, bile and joint fluid.

3. The method of claim 2 wherein the biological sample is urine.

4. The method of claim 1 wherein said measuring step is an immunoassay.

5. The method of claim 4 wherein the immunoassay is performed following hydrolyzation of the sample.

6. The method of claim 1 wherein said measuring step for glucuronidated forms of isoprostanes includes the steps of

separating utilizing column chromatography the free form from glucuronidated forms resulting in a free fraction and glucuronidated fraction,
hydrolyzing the glucuronidated fraction to release the free form and
measuring the released free form as a means of determining the amount of glucuronidated isoprostane.

7. A method to assess oxidative stress in vivo by measuring the amount of glucuronidated isoprostanes in a biological sample containing the isoprostanes and determining the amount of glucuronidated isoprostane present in the sample, and

comparing the measured amount with a control sample and thereby assessing the oxidative stress through said comparison wherein the total amount of glucuronidated isoprostanes increases in the sample from an organism undergoing oxidative stress compared to the control.

8. The method of claim 7 wherein said measuring step is an immunoassay.

9. The method of claim 6 wherein the biological sample is selected from the group consisting of plasma, urine, cerebrospinal fluid, bile and joint fluid.

10. The method of claim 9 wherein the biological sample is urine.

Referenced Cited
U.S. Patent Documents
5700654 December 23, 1997 Roberts et al.
Other references
  • Awad et al: Hepatology, vol. 22(3), pp. 962-968, (1995) (Abstract).
Patent History
Patent number: 5891622
Type: Grant
Filed: Feb 24, 1998
Date of Patent: Apr 6, 1999
Assignees: Oxford Biomedical Research, Inc. (Oxford, MI), Vanderbilt University (Nashville, TN)
Inventors: Jason D. Morrow (Nashville, TN), Hyesook Kim (Bloomfield Hills, MI), L. Jackson Roberts, II (Nashville, TN), Denis M. Callewaert (Oxford, MI)
Primary Examiner: Louise N. Leary
Law Firm: Kohn & Associates
Application Number: 0/28,543